Objective To evaluate the significance and expression of the GDNF/GFRα1/RET genes in distal rectum with Congenital Anorectal Malformations(ARMs) Methods Specimens were collected from resected colon which is 3 cm away to the anus and the distal of rectum in 12 ARMs patients. Haematoxylin and Eosin (HE) staining, immunohistochemical (IHC) staining and reverse transcription PCR (RT-PCR) were used to test RET, GDNF and GFRα1 expression in the differ-ent site of samples with ARMs. Results Expression of ganglia and RET, GDNF and GFRα1 were all negative in distal rec-tum in 12 ARMs patients. Ganglionic cells were found in tissues 3 cm away from the anus where RET, GDNF and GFRα1 expression were also positive in those ARMs patients. Expression of RET, GDNF and GFRα1 were strong positive in middle and low imperforate anus indicated by brown color and week positive in high imperforate anus indicated by yellow color. One case of faeces contamination and one case of loose motion without anal incontinence were found in post-op follow up of high ARMs patients. By contrast, one case of rectal mucosa prolapse and one case of occasional faeces contamination which recov-ered with hip bath were found in post op follow up in middle or low ARMs. Conclusion The unsatisfactory anorectal func-tion is possibly related to the decrease or lost of neurotrophic factors (GDNF/GFRα1/RET) and ganglia in the myenteric plex-uses post plastic surgery in ARMs patients.

Objective To explore the role of trypsinogen activation peptide( TAP)in the early di-agnosis of severe acute pancreatitis( AP)and the prediction of outcome. Methods Eighty-nine cases with pancreatitis were collected containing 45 cases of severe AP and 44 of mild AP. 32 cases of non-AP acute abdominal disease were chosen as control group. Blood of all patients was collected within 6h to measure TAP. ROC curve was used to analyze the results. Results The TAP was different among the three groups (p<0. 01);the TAP of patients with severe AP was the highest. ROC curve indicated that 2. 78 nmol/L was the best cutoff to define AP and non-AP;the sensitivity,specificity and accuracy was 88. 8%,100%and 91. 5%,respectively. 8. 55 nmol/L was used to define severe AP and mild AP;the sensitivity,speci-ficity and accuracy was 92. 9%,95. 2% and 88. 7%,respectively. 11. 20 nmol/L was used to predict the occurrence of complications;the sensitivity,specificity and accuracy was 75. 0%,90. 4% and 82. 2%,re-spectively. Conclusion The TAP can be used to diagnose AP,especially to identify severe AP. The TAP is also beneficial for predicting the outcome of patients with severe AP.

Objective To investigate the relationship between the methylation of RET (proto-oncogene, RET) and Hirschsprung disease (HD), and understand its significance in the development of intestinal wall ganglion cells. Methods Twenty-one surgical removal specimens, which were all dilation segment of HD in Tianjin Children’s Hospital, were used as experimental group, and 5 samples of non-HD normal colon tissues were used as control group. The bisulfite sequencing (BSP)-direct detecting method was used to detect RET CpG island methylation status. The expression of RET protein was detected by immunohistochemistry in experimental group and control group. Results In the experimental group 12 cases (57.14%) were found methylation, but no methylation was found in control group. The average optical density of methylated RET protein was 0.201±0.015 in 12 cases. The average optical density of un-methylated RET protein was 0.364±0.023 in 9 cases (P<0.05). Conclusion RET CpG island methylation reduced protein expression levels of RET. The corollary RET gene methylation may influence the expression levels of RET protein, thereby affecting the ganglion cell development, and thus participating in the occurrence of HD.

The early screening and early diagnosis of biliary atresia is the key to improve the 5-year native liver survival rate,jaundice-free-native-liver survival rate and overall survival rate.So far,the neonatal jaundice,especially that caused by surgical cholestasis exists the lack of awareness.Because of this reason,the delay depletion jaundice patients did not receive early diagnosis and treatment until the severe complications happened.So,it has often missed the best period of operation for the biliary atresia.Called for local medical health governments to formulate the effective early screening methods of biliary atresia and the reasonable schedule of postnatal follow-up,and improving the ability of basic unit medical staff in well understanding cholestasis disease in order to achieve early diagnosis and treatment.Consequently,it will improve the situation of diagnosis and treatment of biliary atresia in our country.

Myeloid-derived suppressor cells (MDSCs) and regulatory T cells (Tregs) are important inhibitory immune cells.Research finds that MDSCs and Tregs increase in a large number under the conditions of inflammation,infection and tumor and they are able to suppress the immunity system and promote tumorigenesis and metastasis through multiple mechanisms.Finding the causes resulting in the increase of MDSCs and Tregs in tumor microenvironment and the eliminating methods have become new focuses in the field of antitumor immunotherapy.

Objective To construct expression vectors of human ribosomal protein S 3(RPS3) and RPS3Ser209 mutant in orcler to investigate the effect of RPS3Ser209 mutant on NF-κB signaling pathway and DNA binding capacity .Methods The vector RPS3-myc was amplified by polymerase chain reaction ( PCR) from the human liver cDNA and subcloned into pcDNA-3.1myc-HisB.RPS3S209A represented mutant RPS3 expression vectors, in which the designated amino acid was mutated to an alanine residue .Dual luciferase reporter gene assay was used to detect the NF-κB transcription activity in HEK293 cells,immunofluorescence to detect RPS3 location, and EMSA to examine NF-κB DNA-binding activity.Results The expression vectors of RPS3-myc and RPS3S209A-myc were constructed.Compared with wild-type RPS3,the nucleus translocation, transactivation activity of NF-κB and DNA binding ability of RPS3S290A were reduced significantly .Conclu-sion The impact of RPS3 on NF-κB signaling pathway depend on its serine 209.

To investigate the prevalence and gene types of KPC in Enterobacteriaceae strains isolated from 4 tertiary general hospitals in Hainan area ,a total 43 isolates which were resistant or intermediate to imipenem or ertapenem were collected from sterile sites between August 2012 and June 2013 from 4 tertiary general hospitals in Hainan area .Modified Hodge Tests (M HT ) were performed for KPC phenotype screening .PCR amplification and DNA sequence were performed to analyze the encoding genes of KPC .Results showed that in the 43 isolates ,21 strains were positive in M HT .PCR and DNA sequence analysis confirmed that 3 isolates produced KPC‐2 .It's suggested that there were the Enterobacteriaceae carrying KPC in Hain‐an area .The encoding genes were KPC‐2 .The KPC gene could be horizontally transmitted by plasmid among different groups of bacteria .It is important to control the transmission of these Enterobacteriaceae carrying KPC .

Objective Acute liver failure is one of the significant causes of death clinically.It is important to explore new serum markers of liver injury for early diagnosis and prognosis prediction of severe liver disease.Hepassocin ( HPS) is a liver-specific mitogenic growth factor.Our study is intended to investigate the correlation between HPS serum levels and the degree of liver injury.Methods Firstly, a mouse model of acute liver injury was constructed via intraperitoneally injection with different doses of CCl4 .Then the survival rate, alanine aminotransferase ( ALT) , aspartate aminotransferase ( AST) and the pathological changes in the liver were detected.Meanwhile, ELISA assay was performed to detect the serum level of HPS.In addition, the mRNA level and protein level of HPS were detected by real-time PCR and Western blotting, respectively.Results The mortality rate was increased and the liver damage was aggravated with the increase of the CCl4 dose.Besides, the ALT and AST levels were also increased in a dose-dependent manner.Additionally, the mRNA and protein levels of HPS were significantly up-regulated and closely related to the degree of the liver injury in the model. Conclusion HPS can be used as a new marker of liver injury in mice.

This study was aimed to screen candidate genes involved in the triterpenoid saponins biosynthetic pathway of the Ilex asprella root. The Illumina platform was applied to perform transcriptomic sequencing of I. asprella root, followed by a series of bioinformatics analysis. The results showed that a total of 272 candidate unigenes were anno-tated to be involved in the biosynthetic pathway of terpenoid in the transcriptome of I. asprella root, including 72 u-nigenes for the upstream pathway and 26 unigenes for cyclization, oxidation and glycosylation in the downstream pathway. Phylogenetic analysis was carried out to further analyze the evolution relationship of some candidate uni-genes and their homologous genes. Two genes IaA S1 and IaA S2 were proved to be mixed amyrin synthases in yeast expression system. Moreover, IaA S1 was identified to one of the rare ASs with α-amyrin as the major product. It was concluded that a series of candidate genes, which might be involved in the biosynthetic pathway of triterpenoid saponins, were screened out from the transcriptome of I. asprella root. Further investigation of these candidate genes will provide insight into their actual functions in the triterpenoid saponins biosynthetic pathway in I. asprella.

Objective To explore the relation between clinical-pathological features,Siewert classification and prognosis of esophagogastric junction (EGJ) carcinoma,and to assess the applicability of the new edition of American Joint Committee of Cancer (AJCC) staging guideline on EGJ adenocarcinoma in China.Methods From 2002 to 2012,the clinical data,pathological features,treatment and prognosis of 218 patients with EGJ malignant tumor were retrospectively analyzed.The patients were typed according to Siewert classification criteria and each case was staged according to 7th edition of AJCC TNM staging criteria for esophagus adenocarcinoma and gastric cancer.Kaplan-Meier method and Log-rank test were performed for survival analysis.Results According to the Siewert classification,type Ⅰ was rare (nine cases,4.1％),type Ⅱ was the most common type (150 cases,68.8％) and followed by type Ⅲ (59 cases,27.1％).There was no significant difference in survival curve among the three types (P＞0.05).The survival curve was drawn according to 7th edition of AJCC TNM staging criteria for esophagus adenocarcinoma.In T staging,the prognosis of patients at T4b was better than that of patients at T4a,the prognosis of patients at ⅡB was better than that of patients at ⅡA.The survival curve of patients at Ⅲ C obviously crossed with that of patients at Ⅳ,which was not in conformity with clinical results.The survival curve was drawn according to 7th edition of AJCC staging criteria for gastric cancer.In T staging,the survival curve of patients at Tis was overlapped with that of patients at T1a.The survival rate of patients at ⅡB could not be accurately predicted by the overall staging.In general,the survival of patients with EGJ carcinoma was better predicted according to 7th edition of AJCC staging criteria for gastric cancer than 7th edition for esophagus adenocarcinoma.Conclusions Neither 7th edition of AJCC staging criteria for esophagus adenocarcinoma nor for gastric cancer could accurately predict its prognosis.In our country,EGJ malignant tumor was similar to gastric cancer and had specific clinical-pathological features.It is necessary to research and establish EGJ carcinoma staging criteria instead of applying the current staging criteria for esophagus adenocarcinoma or gastric cancer.